Application of electrospray mass spectrometry to the structural determination of glycosylphosphatidylinositol membrane anchors. / Nett, Isabelle R. E.; Mehlert, Angela; Lamont, Douglas; Ferguson, Michael A. J.
In: Glycobiology, Vol. 20, No. 5, 05.2010, p. 576-585.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Application of electrospray mass spectrometry to the structural determination of glycosylphosphatidylinositol membrane anchors
A1 - Nett,Isabelle R. E.
A1 - Mehlert,Angela
A1 - Lamont,Douglas
A1 - Ferguson,Michael A. J.
AU - Nett,Isabelle R. E.
AU - Mehlert,Angela
AU - Lamont,Douglas
AU - Ferguson,Michael A. J.
PY - 2010/5
Y1 - 2010/5
N2 - <p>The addition of glycosylphosphatidylinositol (GPI) anchors to proteins is an important posttranslational modification in eukaryotic cells. The complete structural elucidation of GPI anchors is a complex process that requires relatively large amounts of starting material. In this paper, we assess the degree of structural information that can be obtained by applying electrospray mass spectrometry and tandem mass spectrometry to permethylated GPI glycans prepared from a well-characterized GPI-anchored glycoprotein, the variant surface glycoprotein from Trypanosoma brucei. All GPI glycans contain a non-N-acetylated glucosamine residue, and permethylation leads to the formation of a fixed positive charge on the glycans, in the form of a quaternary amine. The permethylated glycans were detected as [M +- Na](2+-) ions, and tandem mass spectrometry of these ions produced substantial, albeit incomplete, structural information on the branching patterns and linkage types for various GPI glycoforms of the variant surface glycoprotein.</p>
AB - <p>The addition of glycosylphosphatidylinositol (GPI) anchors to proteins is an important posttranslational modification in eukaryotic cells. The complete structural elucidation of GPI anchors is a complex process that requires relatively large amounts of starting material. In this paper, we assess the degree of structural information that can be obtained by applying electrospray mass spectrometry and tandem mass spectrometry to permethylated GPI glycans prepared from a well-characterized GPI-anchored glycoprotein, the variant surface glycoprotein from Trypanosoma brucei. All GPI glycans contain a non-N-acetylated glucosamine residue, and permethylation leads to the formation of a fixed positive charge on the glycans, in the form of a quaternary amine. The permethylated glycans were detected as [M +- Na](2+-) ions, and tandem mass spectrometry of these ions produced substantial, albeit incomplete, structural information on the branching patterns and linkage types for various GPI glycoforms of the variant surface glycoprotein.</p>
KW - glycosylphosphatidylinositol
KW - GPI anchor
KW - mass spectrometry
KW - Trypanosoma brucei
KW - variant surface glycoprotein
KW - VARIANT SURFACE GLYCOPROTEIN
KW - TRYPANOSOMA-BRUCEI
KW - PROTEIN
KW - MS
U2 - 10.1093/glycob/cwq007
DO - 10.1093/glycob/cwq007
M1 - Article
JO - Glycobiology
JF - Glycobiology
SN - 0959-6658
IS - 5
VL - 20
SP - 576
EP - 585
ER -